Title of the Study: An open-label, randomized, single-dose, two-treatment, two-sequence, two-way crossover bioequivalence study of Pramipexole prolonged-release tablet Aristo 1.05 mg, manufactured by Advance Pharma GmbH, Germany (Test) with Sifrol 1.05 mg Retardtabletten (prolonged-release 1.05 mg oral tablets), manufactured by Boehringer Ingelheim Pharma GmbH & Co. KG, Germany (Reference), with at least 07 days washout period between each administration, in healthy, adult, male, human subjects under fasting condition.

Study No.: LBS-014-12

Date of Final Report: 29-Jan-13

Principal Investigator

Dr. Mukund Zarapkar

Analytical Investigator

Dr. Ojitkumar Lukram

Study Center:

LifeSan Clinical Research

Address:

Centaur House, Near Hotel Grand Hyatt, Santacruz (East), Mumbai – 400055, India.

Phone: +91-22-66499242

Fax: +91-22-66499239

Total Duration of the Clinical Part:

Stage I – 11 days

Stage II – 11 days

Phase: Bioequivalence study

Objectives:

Primary Objective

To evaluate, whether there exists any bioequivalence between two formulations i.e. Pramipexole prolonged-release tablet Aristo 1.05 mg, manufactured by Advance Pharma GmbH, Germany (Test) with Sifrol 1.05 mg Retardtabletten (prolonged-release 1.05 mg oral tablets), manufactured by Boehringer Ingelheim Pharma GmbH & Co. KG, Germany  (Reference) following single dose administration of one tablet of test or reference formulation in healthy, adult, male, human subjects under fasting condition with at least 07 days washout period by means of rate and extent of absorption based on plasma drug levels of pramipexole.

Secondary Objective

To monitor the safety of the participating subjects in this bioequivalence study of Pramipexole prolonged-release tablet Aristo 1.05 mg, manufactured by Advance Pharma GmbH, Germany (Test) with Sifrol 1.05 mg Retardtabletten (prolonged-release 1.05 mg oral tablets), manufactured by Boehringer Ingelheim Pharma GmbH & Co. KG, Germany  (Reference) determined by means of clinical biochemistry, physical examination and AE/SAE monitoring.

Study Design

·      Open-label

·      Randomized, single dose.

·      Two-treatment, two-sequence, two-way crossover design with a washout period of at least 07 days using two-stage design

·      Bioequivalence study.

·      In healthy, adult, male, human subjects under fasting condition.

Planned for Completion:

46 (18 + 28)

Enrolled and Randomized:

18 (Stage-I) & 28 (Stage-II)

Dropouts:

00

Withdrawals:

02

Completed as Per Protocol:

44

Analyzed:

46

Subjected to Pharmacokinetic and Statistical Evaluation:

44 subjects who completed both periods

Diagnosis and Main Criteria for Inclusion

·      Sex: male.

·      Age: 18–45 years.

·      Healthy, as justified by normal or clinically insignificant findings in clinical and laboratory investigations.

·      BMI: 18.5–30.0 kg/m².

·      Informed consent given in written form.

Test Formulation

Treatment ID:

T

Name:

Pramipexole prolonged-release tablets Aristo 1.05 mg

Strength:

1.05 mg

Batch number:

12518591

Mfg. date:

N/AP

Exp. date:

Jan-2013 [Stage I]

Apr-2013 [Stage II]

Manufacturer:

Dosage form:

Tablets

Dose administered:

One tablet with 240 mL of water

Route of administration:

Oral

Reference Formulation

Treatment ID:

R

Name:

Sifrol retard 1.05 mg Retardtabletten

Strength:

1.05 mg

Lot number:

106692

Mfg. date:

N/AV

Exp. date:

Sep-2014

Manufactured by:

Boehringer Ingelheim Pharma GmbH & Co. KG, Germany

Dosage form:

Tablets

Dose administered:

One tablet with 240 mL of water

Route of administration:

Oral

IMP Administration

Period I

25-Sep-12 (Stage-I) and

28-Nov-12 (Stage-II)

Period II

02-Oct-12 (Stage-I) and

05-Dec-12 (Stage-II)

Duration of Treatments

Each subject received, in a randomized manner and under fasting condition in each period of the study, one tablet of the test formulation (T) or one tablet of reference formulation (R) with a washout period of at least 07 days between the two study periods.

Time Points

0.00 (predose), 1.00, 2.00, 3.00, 4.00, 4.50, 5.00, 5.50, 6.00, 6.50, 7.00, 7.50, 8.00, 8.50, 9.00, 9.50, 10.00, 12.00, 16.00, 24.00, 48.00 and 72.00 hours post dose.

Analytical Methods

LC-MS/MS

LLOQ: 20.0 pg/ml (Stage-I & II)

ULOQ: 3996.3 pg/ml (Stage-I) and 3996.6 pg/ml (Stage-II)

Linearity range: 

For pramipexole – 20.0pg/ml to 3998.7 pg/ml

Criteria for Evaluation

Pharmacokinetics:

Analyzed: Pramipexole

Primary target parameters: Log-transformed C_max andAUC_0-t

Secondary target parameters: t_max, K_el, t_1/2, and AUC_0-inf.

Additional target parameters: MRT, AUC_extrap% and F_rel

Safety Parameter

Clinical laboratory investigations were performed at the time of screening and post-study safety evaluation.

Vital examinations were performed at the time of admission, discharge, on scheduled vital time points and at post-study safety assessment.

AE/SAE/SUSAR profiles of both test and reference formulations were recorded.

Statistical Methods

ANOVA

The primary pharmacokinetic parameters after logarithmic transformation were subjected to an analysis of variance (ANOVA) using the SAS GLM procedure.

To test the SEQUENCE effect the initial model contained the factors SEQUENCE, SUBJECT nested within SEQUENCE, PERIOD and FORM (drug formulation). The significance of the SEQUENCE effect was tested using the SUBJECT nested within SEQUENCE as the error term applying a 5% level of significance. The significance of the PERIOD and FORM effect were tested with a 5% level of significance.

For the estimation of the least squares means (LSM) for the factor FORM a simplified ANOVA was used comprising of SUBJECT, PERIOD and FORM.

For the analysis of the combined data from the two stages, the factor STAGE was included in the ANOVA model.

The test for normality of the residual distribution were performed on Ln-transformed data using the Shapiro-Wilk procedure using a 5% level of significance.

Because of a two-stage study design an a-adjustment of the significance level had to be done with the method of Pocock. For the calculation of the confidence intervals a = 0.0294 instead of a = 0.05 for the interim and the final statistical analysis had to be used, therefore a 94.12% confidence interval had to be calculated instead of a 90% confidence interval.

Consistent with the two one-sided test for bioequivalence, a-adjusted confidence intervals for the difference between drug formulations least-squares means (LSM) of pramipexole were calculated for the log-transformed parameters AUC_0-t, AUC_0-∞ and C_max.

The acceptance range for the a-adjusted confidence band for the ratio of the back-transformed LSMEANS of pramipexole is 80% to 125 % for the parameter AUC_0-t and C_max.

To be inside the acceptance interval the lower bound had to be ≥ 80.00% when rounded to two decimal places and the upper bound had to be ≤ 125.00% when rounded to two decimal places.

In a first step, it was planned to collect data from at least 18 evaluable subjects. If bioequivalence was achieved at stage 1 (interim analysis) the study had to be stopped.

If bioequivalence was not achieved at stage 1 the decision of stopping the study or of continuation of the study with stage 2 had to be based on the sample size re-estimation considering the statistical procedures described in literature.

Because t_max is a discrete variable it is likely that many equal values occur preventing a meaningful analysis the parameter t_max had to be evaluated using the non-parametric Wilcoxon-Mann-Whitney-test.

Confidence Interval [CI]:

The acceptance range for the a-adjusted confidence band for the ratio of the back-transformed LSMEANS of pramipexole had to be 80% to 125 % for the parameter AUC_0-t and C_max.

To be inside the acceptance interval the lower bound had to be ≥ 80.00% when rounded to two decimal places and the upper bound had to be ≤ 125.00% when rounded to two decimal places.

Acceptance criteria:

LnC_max: 80-125% (α-adjusted)

LnAUC_0-t: 80-125% (α-adjusted)

Results

Pharmacokinetics:

Statistical analysis:

The bioequivalence evaluation was based on the log-transformed values of C_max and AUC_0-t ratios (test vs. reference formulation) of pramipexole.

The ratios of the estimated least square means (and 94.12% CIs) of the test to reference formulation (TR) of pramipexole were 95.38% (89.22-101.96%) for LnC_max and 93.04% (84.86-102.00%) for LnAUC_0-t respectively.

The intra-subject CVs for pramipexole for LnC_max, LnAUC_0-t and LnAUC_0-inf were 16.19%, 22.45%, and 22.15%, respectively.

The a-adjusted confidence and for the ratio of the back-transformed LSMEANS of pramipexole (test/reference) for the primary target parameters of C_max and AUC_0-t were within the acceptance interval of 80–125%, thus establishing bioequivalence as per the CPMP note for guidance for Investigation of Bioavailability and Bioequivalence (CPMP/EWP/QWP/1401/98, REV1, CORR. COMMITTEE, January 2010).

Safety Assessment:

During the clinical conduct of the study, twenty-nine (n=29) AE [12 AE in stage-I and 17 AE in stage-II] were recorded, irrespective of the nature of the treatment administered [test or reference]. The subjects were treated appropriately for the AE and it was seen that all the AEs were recovered without any sequel.

Two subjects [S-2 in stage-I & S-39 in stage-II] were withdrawn from the study on account of AE and concomitant medication.

Four more (n=4) AE (2 AEs in stage-I and 2 AE in stage-II) were noted in the clinical laboratory values of the post-study safety assessment.

Conclusion

·      The study was performed according to the protocol and the ICH-GCP guidelines.

·      As per the regulations of “CPMP/EWP/QWP/1401/98, REV.1, CORR. COMMITTEE, January 2010” the a-adjusted confidence band for the ratio of the back-transformed LSMEANS of AUC_0-t and C_max for pramipexole were within the acceptance limits of 80–125%. Hence, it can be concluded that the “Test” formulation, i.e. Pramipexole prolonged-release tablet Aristo 1.05 mg, manufactured by Advance Pharma GmbH, Germany is bioequivalent with the reference formulation i.e. Sifrol 1.05 mg Retardtabletten (prolonged-release 1.05 mg oral tablets) manufactured by Boehringer Ingelheim Pharma GmbH & Co. KG, Germany in terms of the primary target parameters of C_max and AUC_0-t following single dose administration in fasting condition.

·      There were thirty-three [33] adverse events (29 in clinical conduct and 4 in post-study safety assessment), which were managed appropriately till complete resolution without any sequel.

·      Two subjects [S-2 in stage-I & S-39 in stage-II] were withdrawn from the study on account of AE and concomitant medication.

·      Since this study was intended to assess the bioequivalence between test and reference formulations along with the safety of participating the subjects; no statistical comparison of the adverse event profile between test and reference formulation was considered necessary. Apparently, in 13 out of 14 adverse events happened after consuming the reference formulation had some relationship determined with the pramipexole, whereas relationship with Pramipexole could be concluded in 14 out of 15 adverse events that happened after consuming the test formulation. There was no relationship of two AE with the study drug [AE-12 – sequence TR in period II of stage-I was not related with reference formulation & AE-10 – sequence TR in period II of stage-II which was not related with test formulation]. All the adverse events noted in this study were in accordance with the SmPC of Sifrol 1.05 mg Retardtabletten (prolonged-release 1.05 mg oral tablets) and therefore, no new safety related finding could be elicited.